|Table of Contents|

Development of hybridoma cell lines that secrete monoclonal antibodies against diethyl phthalate

Chinese Journal of Applied & Environmental Biology[ISSN:1006-687X/CN:51-1482/Q]

2019 03
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Development of hybridoma cell lines that secrete monoclonal antibodies against diethyl phthalate
ZHOU YongZHANG JiaqiLIAO HaoxiangMA Yuning** & CHENG Jinping
School of Environmental Science and Engineering, Shanghai Jiaotong University, Shanghai 200240, China
diethyl phthalate (DEP) hybridoma cell line McAb HAT screening colloidal gold immunochromatographic (GICA)

To establish a rapid immunological method for the detection of diethyl phthalate (DEP) and other phthalic acid esters (PAEs), highly efficient and stable hybridoma cell lines that secrete anti-DEP monoclonal antibodies (McAb) are urgently needed. In this study, mepridine-bovine serum albumin (MEP-BSA) was prepared, its quality was confirmed by mass spectrometry, and then it was used as an immune antigen to immunize BALb/c mice so that serum titers in them could be determined by an indirect competitive enzyme-linked immunosorbent assay (ELISA). The technique of cell hybridoma screening was then used to screen for hybridoma cell lines resistant to anti-DEP monoclonal antibodies. Results showed that MEP-BSA was successfully synthesized. The mice’s serum titer reached a maximum value of 1:5000, and 17 cell lines that could secrete anti-DEP monoclonal antibodies were identified after selection using HAT (hypoxanthine-aminopterin-thymidine). Subclones of these were then prepared whose cell supernatant titers could reach a maximum value of 1:10000. The successfully constructed hybridoma cell lines could stably secrete anti-DEP monoclonal antibodies, resulting in an inhibition ratio as high as 86.5%, and the cross-reaction rate of the 11 structural analogs obtained was less than 0.1%. Our method provides a stable and efficient source of monoclonal antibodies for use in ELISA-based detection methods, as well as a reference for the development of the colloidal gold immunochromatographic (GICA) method for the ELISA detection of DEP GICA strips.


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Last Update: 2019-06-25